Investigation of mechanism of action of Kuhuang Granules against intrahepatic cholestasis through the integration of network pharmacology and animal experiments

Huang Juan; Fang Wei; Xiao Yaping; Du Biao

doi:10.13406/j.cnki.cyxb.003592

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Investigation of mechanism of action of Kuhuang Granules against intrahepatic cholestasis through the integration of network pharmacology and animal experiments

Basic research | 更新时间：2025-10-21

- Investigation of mechanism of action of Kuhuang Granules against intrahepatic cholestasis through the integration of network pharmacology and animal experiments
- Journal of Chongqing Medical University Vol. 49, Issue 9, Pages: 1113-1120(2024)
- 作者机构：
  
  1.川北医学院药学院，南充 637000
  2.重庆大学附属三峡医院药学部，重庆 404000
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.003592
  CLC： R285.5
- Received：30 November 2023，
  
  Revised：2024-01-14，
  
  Accepted：18 January 2024，
  
  Published：28 September 2024
- 稿件说明：
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黄娟,方伟,肖亚平,等.基于网络药理学和动物实验探讨苦黄颗粒抗肝内胆汁淤积的作用机制[J].重庆医科大学学报,2024,49(9):1113-1120.

Huang Juan,Fang Wei,Xiao Yaping,et al.Investigation of mechanism of action of Kuhuang Granules against intrahepatic cholestasis through the integration of network pharmacology and animal experiments[J].Journal of Chongqing Medical University,2024,49(9):1113-1120.
黄娟,方伟,肖亚平,等.基于网络药理学和动物实验探讨苦黄颗粒抗肝内胆汁淤积的作用机制[J].重庆医科大学学报,2024,49(9):1113-1120. DOI： 10.13406/j.cnki.cyxb.003592.

Huang Juan,Fang Wei,Xiao Yaping,et al.Investigation of mechanism of action of Kuhuang Granules against intrahepatic cholestasis through the integration of network pharmacology and animal experiments[J].Journal of Chongqing Medical University,2024,49(9):1113-1120. DOI： 10.13406/j.cnki.cyxb.003592.

摘要

目的

通过整合动物实验和网络药理学的方法，探讨苦黄颗粒抗肝内胆汁淤积的潜在作用机制。

方法

评估苦黄颗粒对α-萘异硫氰酸酯（alpha-naphthyl isothiocyanate，ANIT）诱导的胆汁淤积性肝损伤小鼠的保肝作用，考察肝体比、血清学指标、氧化应激等生化指标及肝脏病理特征。结合网络药理学和生物信息学挖掘苦黄颗粒治疗肝内胆汁淤积的潜在靶点和关键通路。通过western blot验证相关关键通路。

结果

动物实验表明，与空白组相比，模型组肝体比、碱性磷酸酶（alkaline phosphatase，AKP）、丙氨酸转氨酶（alanine transaminase，ALT）、天冬氨酸转氨酶（aspartate aminotransferase，AST）、直接胆红素（direct bilirubin，DBIL）、总胆红素（total bilirubin，TBIL）、总胆汁酸（total bile acid，TBA）、丙二醛（malondialdehyde，MDA）、胆固醇7α-羟化酶（cholesterol 7α-hydroxylase，CYP7A1）蛋白表达水平均升高（

0.05）；而体质量、超氧化物歧化酶（super oxide dismutase，SOD）、谷胱甘肽-过氧化氢酶（glutathione peroxidase，GSH-Px）、法尼醇X受体（farnesoid X receptor，FXR）、小异二聚体伴侣（small heterodimer partner，SHP）抗体、胆盐输出泵（bile salt export pump，BSEP）和多药耐药相关蛋白2（multidrug resistance-

associated protein 2，MRP2）蛋白表达水平均降低（

0.05）。与模型组比较，苦黄颗粒各剂量组和联苯双酯组肝体比、TBIL、DBIL、TBA、ALT、AST、AKP、MDA、CYP7A1的水平均下降（

0.05）；同时体质量、SOD、GSH-Px、FXR、SHP、BSEP和MRP2升高（

0.05）。网络药理学分析表明，苦黄颗粒可能通过调控肿瘤坏死因子（tumor necrosis factor，TNF）、核因子-κB1（nclear factor kappa b subunit1，NF-κB1）、法尼醇X受体（farnesoid x receptor，FXR）和药耐受性相关蛋白2（multidrug resistance-associated protein 2，MRP2）等靶点影响胆汁酸的分泌减轻胆汁淤积肝损伤。

结论

苦黄颗粒可能通过激活FXR通路相关靶点，减少胆汁酸合成，促进胆汁酸排泄，从而减轻胆汁淤积肝损伤。

Abstract

Objective

To elucidate the potential mechanism of action of Kuhuang Granules against intrahepatic cholestasis by employing a comprehensive approach that combines animal experiments with network pharmacology.

Methods

We assessed the hepatoprotective effect of Kuhuang Granules in mice with cholestatic liver injury induced by alpha-naphthyl isothiocyanate by evaluating the liver-to-body ratio，serological indicators，oxidative stress，and other blood biochemical parameters，and pathological characteristics of the liver. The potential targets and key pathways of Kuhuang Granules in the treatment of intrahepatic cholestasis were uncovered through the integration of network pharmacology and bioinformatics. The identified key pathways were validated through Western blot analysis.

Results

Animal experiments showed that，compared with the blank group，the model group exhibited significantly increased levels of liver-to-body ratio，alkaline phosphatase（AKP），alanine transaminase（ALT），aspartate aminotransferase（AST），direct bilirubin（DBil），total bilirubin（TBil），total bile acid（TBA），malondialdehyde（MDA），and cholesterol 7α-hydroxylase（CYP7A1） protein expression（all

0.05）；the body mass，superoxide dismutase （SOD），glutathione peroxidase（GSH-Px），farnesoid X receptor（FXR），small heterodimer partner（SHP） antibodies，bile salt export pump（BSEP），and mult

idrug resistance-associated protein 2（MRP2） expression levels were significantly decreased（all

0.05）. Compared with the model group，the Kuhuang Granules groups and bifendate group showed significantly decreased levels of liver-to-body ratio，TBil，DBil，TBA，ALT，AST，AKP，MDA，and CYP7A1（all

0.05）； meanwhile，the body mass，SOD，GSH-Px，FXR，SHP，BSEP，and MRP2 were significantly increased（all

0.05）. Network pharmacology analysis indicated that Kuhuang Granules may alleviate cholestatic liver injury by affecting bile acid secretion through regulating targets such as tumor necrosis factor，nuclear factor kappa b subunit 1，FXR，and MRP2.

Conclusion

Kuhuang Granules may alleviate cholestatic liver injury by activating targets associated with the FXR pathway，reducing bile acid synthesis，and enhancing bile acid excretion.

关键词

Keywords

references

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